Protein fixation with glutaraldehyde is required before staining the gel with Coomassie Brilliant Blue dye (PageBlue™ Protein Staining Solution, #R0571) or silver (PageSilver™ Silver Staining Kit, #K0681).
Other compounds commonly used for protein fixation (e.g., acetic acid, isopropanol, ethanol, TCA) are not suitable; proteins will wash away during staining procedure.
- Add 100 ml of deionized or distilled water to the gel and wash for 1 min with gentle agitation. Discard the wash.
- Add 50 ml of freshly prepared 5% glutaraldehyde solution (gel should be covered completely). Fix with gentle agitation for 30 min. Discard the solution.
- Add 100 ml of deionized or distilled water to the gel and wash for 5 minutes with gentle agitation. Discard the wash. Repeat this step twice. The gel is now ready for staining.
For detailed staining protocols using Fermentas PageBlue™ Protein Staining Solution or PageSilver™ Silver Staining Kit, refer to the product manuals.
Proceed directly to the staining step in PageBlue™ Protein Staining Solution protocol and to sensitizing step in PageSilver ™ Silver Staining Kit protocol.